CN104105692A - Benzotriazine oxides as drugs targeting mycobacterium tuberculosis - Google Patents

Benzotriazine oxides as drugs targeting mycobacterium tuberculosis Download PDF

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CN104105692A
CN104105692A CN201280069153.0A CN201280069153A CN104105692A CN 104105692 A CN104105692 A CN 104105692A CN 201280069153 A CN201280069153 A CN 201280069153A CN 104105692 A CN104105692 A CN 104105692A
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P·马德瑞德
S·肖帕
K·莱恩
G·库佩
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Standford Institute for International Studies
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Abstract

Benzotriazine doxides are disclosed as drugs targeting mycobacterium tuberculosis, including novel compounds of formula I: (Formula (I)

Description

As the benzotriazine oxides of mycobacterium tuberculosis targeted drug
Applicant: think to grind (SRI) world, Men Luo Parker city, California (Menlo Park, CA)
Contriver: P Ma Derui moral, Saratoga city, California (Saratoga, CA), United States citizen
Xiao's S handkerchief, mountain scene city, California (Mountain View, CA), citizen of india
K Lay grace, Sen Niweier city, California (Sunnyvale, CA), United States citizen
Wear in G storehouse, Te Leixi city, California (Tracy, CA), United States citizen
This project is by bonus R56AI090817, NIH (National Institutes of Health) and the national allergy of national allergy and Infectious Disease Research Institute (National Institute of Allergy and Infectious Diseases) and supporting with about HHSN266200600011C (N01-AI-60011) and No. P19748l appropriation of/NIAID of Health and Public Service Department (Department of Health and Human Services) of Infectious Disease Research Institute (National Institute of Allergy and Infectious Diseases); Government enjoys certain right to the present invention.
The application requires the right of priority of No. 61567829 U.S. Patent application of submitting on December 7th, 2011.
Brief introduction
Current, research and development (the R & D) channel of new tuberculosis (TB) medicine is not enough to solve the drug resistance strain occurring.Therefore, in the urgent need to filling up early stage drug development channel with slowing down extra drug-fast new treatment.Since nineteen fifty, in the time showing that coupling medicine, particularly Fungicidal compounds have delayed drug-fast generation, combination chemotherapy has been just the nursing standard of TB. 1the next important breakthrough of TB treatment is to comprise Rifampin and pyrazinoic acid amide in multiple medicines mixture (multidrug cocktail), and this is because these medicines are stayed the fungicidal activity of bacterial flora and promote significantly the removing of infecting holding by it.Short distance DOTS (Direct Observed Therapy Short-course, the DOTS) scheme of current recommendation is effective at medicine susceptibility TB camber, but needs the treatment time of at least 6 months.The next important breakthrough of TB treatment is the improved plan that can develop shorter treatment time of a kind of needs, thereby reduces costs, improves patient compliance and slow down the appearance of TB (MDR-TB) bacterial strain of multidrug resistance.
To many medicines have tolerance non-copy to hold stay (nonreplicating persistence, NRP) mycobacterium tuberculosis (the Mycobacterium tuberculosis of stage survival, Mtb), provide possible explanation for eliminating the required long treatment plan of infection. 2the exploitation of new combined treatment, comprises the medicine that these NRP stages of Mtb are had microbe killing properties, and has the potential that shortens treatment plan duration.Metronidazole---a kind of 5-nitroimidazole microbiotic that is mainly used in treating anaerobic bacterium infection---is that NRP Mtb is shown to one of bactericidal linearize compound. 2two kinds of newer 5-nitroimidazoles---PA-8243 and OPC-676834---have stronger effect to Mtb and just by assessing clinically with standard TB drug combination.Up to now, these two kinds of 5-nitroimidazoles have demonstrated and have had early stage fungicidal activity in patient. 5,6the preclinical study of the mouse model of TB shows, in scheme for combining, comprises the germ killing drugs with anti-NRP Mtb activity and shortened and cure the required time of mouse infection, 7but these find the duration that is interpreted as shortening clinical treatment still to await determining.
In order to develop the new drug that shortens TB treatment time, we have assessed the activity of the opposing Mtb H37Rv of the biological reducing activating compounds of the other types of the anti-microbial activity with previous report.We disclose the BTO of the tuberculosis compound new as a class, and these BTO have many special performances, and these unique performances make it be particularly suitable as anti-TB medicine, particularly have the potential that shortens treatment duration.
Pertinent literature comprises: Chopra etc., J Med Chem.2012 Jul 12; 55 (13): 6047-60; Hay, etc., J.Med.Chem., the 2008,51 (21), the 6853-6865 page; Zeman, etc., Int.J.Radiat.Oncol., Biol., Phys.1989,16,977-981; Minchinton, etc., Int.J.Radiat.Oncol., Biol., Phys 1992,22,701-705; Kelson, etc., Anti-Cancer Drug Des.1998,13,575-592; Jiang, etc., Bioorg.Med.Chem.Lett.2006,16,4209-4213; And Jiang, etc., Arch.Pharm. (Weinheim, Ger.) 2007,340,258-263.
Summary of the invention
In one embodiment, the invention provides compound or its pharmacy acceptable salt or the steric isomer with formula I structure:
Wherein: each X is H, halogen, alkyl, OR, SR, NR ' R, BR ' R, heterocycle or another functional group independently, wherein each R is H, halogen, alkyl or another functional group independently; W is N, C, O, S, H or B or another connection atom; Each A and B are H or the optional alkyl replacing, and it can be connected to optionally assorted-cycloalkyl; And Z is 4-8 ring optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring, at 6,7-, 5, condense on 6-or 7,8-position.
In specific embodiments, each X is H, halogen, alkyl, OR, SR, NR ' R, BR ' R independently, and wherein each R is H, halogen or alkyl independently, is more particularly H; W is N, C, O, S or B, is more particularly N; Each A and B are H or the optional alkyl replacing, and it can be connected to optionally assorted-cycloalkyl, are more particularly optional piperidyl or the pyrrolidyl replacing; And Z is 4-8 ring optional existence, saturated or undersaturated, optional replacement, itself and phentriazine ring are 6,7-, 5, condenses on 6-or 7,8-position, be 5 or 6 rings that optionally exist, saturated or undersaturated, optional replacement especially, itself and phentriazine ring condense on 7,8-position, are more particularly that exist, undersaturated 5 rings, itself and phentriazine ring condense on 7,8-position.
In specific embodiments:
Each X is H, halogen, alkyl, OR, SR, NR ' R or BR ' R independently, and wherein each R is H, halogen or alkyl independently; W is N, C, O, S or B; Each A and B are H or the optional alkyl replacing, and it can be connected to optionally assorted-cycloalkyl; And Z is 4-8 ring optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring, at 6,7-, 5, condense on 6-or 7,8-position.
Each X is H, halogen, alkyl, OR, SR, NR ' R or BR ' R independently, and wherein each R is H, halogen or alkyl independently; W is N, C, O, S or B; Each A and B are the optional alkyl replacing, and it can be connected to optionally assorted-cycloalkyl; And Z is 4-8 ring optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring, at 6,7-, condense on 5,6-or 7,8-position.
Each X is H, halogen, alkyl, OR, SR, NR ' R or BR ' R independently, and wherein each R is H, halogen or alkyl independently; W is N; Each A and B are the optional alkyl replacing, and it can be connected to Heterocyclylalkyl; And Z is 5 or 6 rings optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring condense on 7,8-position.
Each X is H; W is N; A and B are connected to piperidyl or the pyrrolidyl of optional replacement; And Z is undersaturated 5 rings, and itself and phentriazine ring condense on 7,8-position.
In specific embodiments, compound is the BTO with structure disclosed herein, that specifically enumerate, as shown in the table.
In other embodiments, the invention provides and comprise topic and state pharmaceutical composition and the test kit of the BTO compound Killing Mycobacterium Tuberculosis different with the second (Mtb) medicine.
The present invention also provides to prepare and has inscribed the method for stating BTO compound, comprises the oxidizing reaction that uses HOF:ACN.
The method that the present invention also provides treatment mycobacterium tuberculosis (Mtb) to infect, comprising: make the people of needs contact titled reference compound or the composition of significant quantity.
On the other hand, the invention provides the method that treatment mycobacterium tuberculosis (Mtb) infects, comprising: make the people of needs contact 1,2 of significant quantity, 4-phentriazine two-N-oxide compound (BTO).
The use that topic is stated or the method for the treatment of can further comprise the subsequent step that detects resulting infection minimizing and/or the preliminary step that detects infection.
In specific embodiments, infect comprise non-copy to hold stay (NRP) Mtb cell.
All combinations that the present invention comprises specific embodiments and preferred embodiment, as every kind of combination all by hardy, enumerate individually.
Brief description of the drawings
Fig. 1. after the oral each compound that gives 100mg/kg, the 8a in female Balb/c mouse (n=3 mouse/time point) and the plasma drug level of 20q.After administration, in 24 hours, gather blood at each time point, blood treatment is become to blood plasma, and each test compounds is analyzed.Two kinds of compounds in the concentration at 24 hours point places lower than quantitative limit.
Embodiment
Following description to specific embodiments and embodiment provides by way of example, instead of provides by the mode of restriction.It will be readily appreciated by those skilled in the art that to change and maybe can revise various non-key parameters, to obtain substantially similar result.Explain unless the contrary indication or separately, in these are described and in the application's full text, term " one " mean more than one, and term "or" means and/or, and polynucleotide sequence is interpreted as comprising opposite strand and alternative skeleton described herein.What in addition, class was all class members writes a Chinese character in simplified form; For example (C1-C3) alkyl is all C1-C3 alkyl: the writing a Chinese character in simplified form of methyl, ethyl and propyl group (comprising its isomer).Term used herein " heteroatoms " means any atom except carbon, hydrogen or oxygen conventionally.Preferred heteroatoms comprises oxygen (O), phosphorus (P), sulphur (S), nitrogen (N), silicon (S), arsenic (As), selenium (Se) and halogen, and preferred heteroatom functional group is halo formyl radical, hydroxyl, aldehyde, amine, azo-group, carboxyl, cyano group, thiocyanogen, carbonyl, halo, hydrogen peroxide-based, imines, aldimine, isonitrile, isocyanate group, itrate group, nitrile, nitrous acid ester group, nitro, nitroso-group, phosphate-based, phosphine carboxylic ester group, sulfide, alkylsulfonyl, sulfenyl and sulfydryl.
Unless otherwise mentioned, term " alkyl ", or itself is as another substituent part, means straight or branched or cyclic hydrocarbon group, or its combination, it is completely saturated, has the carbonatoms (, C1-C8 means 1 to 8 carbon) of appointment.The example of alkyl comprises methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, the tertiary butyl, isobutyl-, sec-butyl, cyclohexyl, (cyclohexyl) methyl, cyclopropyl methyl, such as homologue and the isomer of n-pentyl, n-hexyl, n-heptyl, n-octyl etc.
Term " thiazolinyl ", or itself is as another substituent part, means straight or branched or cyclic hydrocarbon group, or its combination, it can be monounsaturated or polyunsaturated, has the carbonatoms (, C2-C8 means 2 to 8 carbon) of appointment and more than one two keys.The example of thiazolinyl comprises vinyl, 2-propenyl, crot(on)yl, 2-isopentene group, 2-(butadienyl), 2,4-pentadienyl, 3-(Isosorbide-5-Nitrae-pentadienyl) and more senior homologue and isomer thereof.
Term " alkynyl ", or itself is as another substituent part, means straight or branched alkyl, or its combination, it can be monounsaturated or polyunsaturated, has carbonatoms (, C2-C8 means 2 to 8 carbon) and more than one triple bond of appointment.The example of alkynyl comprises ethynyl, 1-proyl and 3-proyl and more senior homologue and isomer thereof.
Term " alkylidene group ", or itself is as another substituent part, means the divalent group derived from alkyl, for example-CH 2-CH 2-CH 2-CH 2-.Conventionally, alkyl (or alkylidene group) has 1 to 24 carbon atom, and in the present invention, these groups preferably have 10 following carbon atoms." low alkyl group " or " low-grade alkylidene " is compared with the alkyl of short chain or alkylidene group, conventionally has 8 following carbon atoms.
Term " alkoxyl group ", " alkylamino " and " alkyl sulfenyl " (or thio alkoxy) use with its conventional sense, and refer to those alkyl that are connected with the remainder of molecule by Sauerstoffatom, amino or sulphur atom respectively.
Unless otherwise mentioned, term " assorted alkyl ", itself or with the combination of another term, mean stable straight or branched or cyclic hydrocarbon group, or its combination, carbon atom and 1 to 3 heteroatoms that is selected from O, N, Si and S by defined amount form, and wherein nitrogen and sulphur atom are optionally oxidized, and nitrogen heteroatom is optionally quaternized.Heteroatoms O, N and S can be placed in any interior location of assorted alkyl.Heteroatoms Si can be placed in the optional position of assorted alkyl, comprises the position that alkyl is connected with the remainder of molecule.Comprise-CH of example 2-CH 2-O-CH 3,-CH 2-CH 2-NH-CH 3,-CH 2-CH 2-N (CH 3)-CH 3,-CH 2-S-CH 2-CH 3,-CH 2-CH 2,-S (O)-CH 3,-CH 2-CH 2-S (O) 2-CH 3,-CH=CH-O-CH 3,-Si (CH 3) 3,-CH 2-CH=N-OCH 3with-CH=CH-N (CH3)-CH 3.Maximum two heteroatomss can be continuous, for example, and-CH 2-NH-OCH 3with-CH 2-O-Si (CH 3) 3.
Similarly, term " assorted alkylidene group ", or itself is as another substituent part, means the divalent group derived from assorted alkyl, for example-CH 2-CH 2-S-CH 2-CH 2-and-CH 2-S-CH 2-CH 2-NH-CH 2-.For assorted alkylidene group, heteroatoms also can occupy one or two end of the chain (for example, alkylidene group oxygen base, alkylenedioxy group, alkylidene amino, alkylidene group diamino etc.).In addition,, for alkylidene group and assorted alkylidene group linking group, do not indicate the orientation of linking group.
Unless otherwise mentioned, term " cycloalkyl " and " Heterocyclylalkyl ", itself or be combined with other terms, the annular form of representative " alkyl " and " mix alkyl " respectively.Therefore, cycloalkyl has the carbonatoms (, C3-C8 means 3 to 8 carbon) of appointment and also can have one or two pair of key.Heterocyclylalkyl is made up of the carbon atom specifying number and 1 to 3 heteroatoms that is selected from O, N, Si and S, and wherein nitrogen and sulphur atom are optionally oxidized and nitrogen heteroatom is optionally quaternized.In addition,, for Heterocyclylalkyl, heteroatoms can occupy the position that heterocycle is connected with the remainder of molecule.The example of cycloalkyl comprises cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, suberyl etc.The example of Heterocyclylalkyl comprises 1-(1,2,5,6-tetrahydro pyridyl), piperidino, 2-piperidyl, 3-piperidyl, 4-morpholinyl, morpholinyl, tetrahydrofuran (THF)-2-base, tetrahydrofuran (THF)-3-base, tetramethylene sulfide-2-base, tetramethylene sulfide-3-base, 1-piperazinyl, 2-piperazinyl etc.
Unless otherwise mentioned, term " halo " and " halogen ", or itself is as another substituent part, means fluorine, chlorine, bromine or iodine atom.In addition, term means to comprise that as " haloalkyl " described halogen atom can be identical or different by 1 alkyl to the individual halogen atom replacement of (2m '+1) (wherein m ' is the sum of carbon atom in alkyl).For example, term " halo (C1-C4) alkyl " means to comprise trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl etc.Therefore, term " haloalkyl " comprises single haloalkyl (alkyl being replaced by a halogen atom) and multi-haloalkyl (by 2 alkyl to (2m '+1) individual halogen atom replacement, wherein m ' is the sum of carbon atom in alkyl).Unless otherwise mentioned, term " whole haloalkyl " means the alkyl being replaced by the individual halogen atom of (2m '+1) (wherein m ' is the sum of carbon atom in alkyl).For example, term " perhalogeno (C1-C4) alkyl " means to comprise trifluoromethyl, pentachloro-ethyl, 1,1, the bromo-2-chloroethyl of the fluoro-2-of 1-tri-etc.
Term " acyl group " refers to by the hydroxylic moiety except disacidify derived from those groups of organic acid.Therefore, acyl group means to comprise, for example, and ethanoyl, propionyl, butyryl radicals, decanoyl, valeryl, benzoyl etc.
Unless otherwise mentioned, that term " aryl " means is polyunsaturated, be generally aromatic hydrocarbon substituent, and it can be and condenses together or covalently bound monocycle or many rings (being up to three rings).The limiting examples of aryl comprises phenyl, 1-naphthyl, 2-naphthyl, 4-xenyl and 1,2,3,4-tetralin.
Term " heteroaryl " refers to and contains 0 to 4 heteroatomic aryl that is selected from N, O and S (or ring), and wherein nitrogen and sulphur atom are optionally oxidized and nitrogen heteroatom is optionally quaternized.Heteroaryl can be connected to by heteroatoms the remainder of molecule.The unrestricted example of heteroaryl comprises 1-pyrryl, 2-pyrryl, 3-pyrryl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purine radicals, 2-benzimidazolyl-, 5-indyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl and 6-quinolyl.
For simplicity, term " aryl ", in the time being combined with other terms, (for example, aryloxy, aryl sulphur oxygen base, arylalkyl), comprises aromatic ring and hetero-aromatic ring as defined above.Therefore, term " arylalkyl " means to comprise that those groups that aryl is connected to alkyl (for example, benzyl, styroyl, pyridylmethyl etc.), described alkyl comprises that carbon atom (for example, methylene radical) for example, by for example alternative those alkyl (, Phenoxymethyl, 2-pyridyl oxygen ylmethyl, 3-(1-naphthyl oxygen base) propyl group etc.) of Sauerstoffatom.
Each above-mentioned term (for example, " alkyl ", " assorted alkyl ", " aryl " and " heteroaryl ") means to comprise the replacement form of described group and does not replace two kinds of forms.The preferred substituents of the group of every type provides as follows.
The substituting group of alkyl and assorted alkyl group (and be called as alkylidene group, thiazolinyl, assorted alkylidene group, assorted thiazolinyl, alkynyl, cycloalkyl, Heterocyclylalkyl, cycloalkenyl group and heterocycloalkenyl those groups) can be to be selected from following various groups :-OR ' ,=O ,=NR ' ,=N-OR ' ,-NR ' R " ,-SR ', halogen ,-SiR ' R " R " ' ,-OC (O) R ' ,-C (O) R ' ,-CO 2r ' ,-CONR ' R " ,-OC (O) NR ' R " ,-NR " C (O) R ' ,-NR '-C (O) NR " R " ' ,-NR '-SO 2nR " ' ,-NR " CO 2r ' ,-NH-C (NH 2)=NH ,-NR ' C (NH 2)=NH ,-NH-C (NH 2)=NR ' ,-S (O) R ' ,-SO 2r ' ,-SO 2nR ' R " ,-NR " SO 2r ,-CN and-NO 2, substituent number is 0 to 3, particularly preferably has 0,1 or 2 substituent those group.R ', R " and R " ' refer to independently of one another hydrogen, unsubstituted (C1-C8) alkyl and assorted alkyl, unsubstituted aryl, the aryl, unsubstituted alkyl, alkoxyl group or the thio alkoxy that are replaced by 1 to 3 halogen, or aryl-(C1-C4) alkyl.As R ' and R, " while being connected to identical nitrogen-atoms, it can be combined with nitrogen-atoms and form 5,6 or 7 rings.For example ,-NR ' R " means to comprise 1-pyrrolidyl and 4-morpholinyl.Conventionally, alkyl or assorted alkyl have 0 to 3 substituting group, and the present invention preferably has two following substituent those groups.More preferably, alkyl or assorted alkyl are unsubstituted or mono-substituted.Most preferably, alkyl or assorted alkyl are unsubstituted.To substituent discussion, it will be understood by those skilled in the art that term " alkyl " means to comprise group (for example ,-CF such as tri haloalkyl from above-mentioned 3with-CH 2cF 3).
The preferred substituents of alkyl and assorted alkyl is selected from :-OR ' ,=O ,-NR ' R " ,-SR ', halogen ,-SiR ' R " R " ' ,-OC (O) R ' ,-C (O) R ' ,-CO 2r ' ,-CONR ' R " ,-OC (O) NR ' R " ,-NR " C (O) R ' ,-NR " CO 2r ' ,-NR '-SO 2nR " R " ' ,-S (O) R ' ,-SO2R ' ,-SO 2nR ' R " ,-NR " SO 2r ,-CN and-NO 2, wherein R ' and R " as above definition.Further preferred substituting group is selected from :-OR ' ,=O ,-NR ' R ", halogen ,-OC (O) R ' ,-CO 2r ' ,-CONR ' R " ,-OC (O) NR ' R " ,-NR " C (O) R ' ,-NR " CO 2r ' ,-NR '-SO 2nR " R " ' ,-SO 2r ' ,-SO 2nR ' R " ,-NR " SO 2r ,-CN and-NO 2.
Similarly, the substituting group of aryl and heteroaryl is various and is selected from: halogen ,-OR ' ,-OC (O) R ' ,-NR ' R " ,-SR ' ,-R ' ,-CN ,-NO 2,-CO 2r ' ,-CONR ' R " ,-C (O) R ' ,-OC (O) NR ' R " ,-NR " C (O) R ' ,-NR " CO 2r ' ,-NR '-C (O) NR " R " ' ,-NR '-SO 2nR " R " ' ,-NH-C (NH2)=NH ,-NR ' C (NH 2)=NH ,-NH-C (NH 2)=NR ' ,-S (O) R ' ,-SO 2r ' ,-SO 2nR ' R " ,-NR " SO 2r ,-N 3,-CH (Ph) 2, perfluor (C1-C4) alkoxyl group and perfluor (C1-C4) alkyl, substituent number is the sum of the 0 open valency to aromatic nucleus system (open valence); And wherein R ', R " and R " ' is independently selected from hydrogen, (C1-C8) alkyl and assorted alkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C1-C4) alkyl and (unsubstituted aryl) oxygen base-(C1-C4) alkyl.In the time that aryl is 1,2,3,4-naphthane, it can be substituted or unsubstituted (C3-C7) spiro cycloalkyl group replaces.(C3-C7) mode that spiro cycloalkyl group can be identical with " cycloalkyl " that limit is herein substituted.Conventionally, aryl or heteroaryl have 0 to 3 substituting group, and the present invention preferably has two following substituent those groups.In one embodiment of the invention, aryl or heteroaryl are unsubstituted or mono-substituted.In another embodiment, aryl or heteroaryl are unsubstituted.
The preferred substituents of aryl and heteroaryl is selected from: halogen ,-OR ' ,-OC (O) R ' ,-NR ' R " ,-SR ' ,-R ' ,-CN ,-NO 2,-CO 2r ' ,-CONR ' R " ,-C (O) R ' ,-OC (O) NR ' R " ,-NR " C (O) R ' ,-S (O) R ' ,-SO 2r ' ,-SO 2nR ' R " ,-NR " SO 2r ,-N 3,-CH (Ph) 2, perfluor (C1-C4) alkoxyl group and perfluor (C1-C4) alkyl, wherein R ' and R " as above definition.Further preferred substituting group is selected from: halogen ,-OR ' ,-OC (O) R ' ,-NR ' R " ,-R ' ,-CN ,-NO 2,-CO 2r ' ,-CONR ' R " ,-NR " C (O) R ' ,-SO 2r ' ,-SO 2nR ' R " ,-NR " SO 2r, perfluor (C1-C4) alkoxyl group and perfluor (C1-C4) alkyl.
Substituting group-CO used herein 2h comprises that its bioisostere replaces (bioisosteric replacement); Referring to, for example, The Practice of Medicinal Chemistry; Wermuth, C.G., Ed.; Academic Press:New York, 1996; The 203rd page.Two substituting groups on the adjacent atom of aromatic ring or hetero-aromatic ring are optionally by formula-T-C (O)-(CH 2) substituting group of q-U-substitutes, wherein T and U are-NH-,-O-,-CH independently 2-or singly-bound, and q is 0 to 2 integer.Or two substituting groups on the adjacent atom of aromatic ring or hetero-aromatic ring are optionally substituted by the substituting group of formula-A-(CH2) r-B-, wherein A and B are-CH independently 2-,-O-,-NH-,-S-,-S (O)-,-S (O) 2-,-S (O) 2nR '-or singly-bound, and r is 1 to 3 integer.One of singly-bound of the new ring forming is optionally substituted by two keys.Or two substituting groups on the adjacent atom of aromatic ring or hetero-aromatic ring are optionally by formula-(CH 2) s-X-(CH 2) t--substituting group substitute, wherein s and t are 0 to 3 integer independently, and X be-O-,-NR '-,-S-,-S (O)-,-S (O) 2-or-S (O) 2nR '.-NR '-and-S (O) 2nR '-in substituent R ' be selected from hydrogen or unsubstituted (C1-C6) alkyl.Term " pharmacy acceptable salt " means to comprise the salt of the active compound of preparing with relatively nontoxic acid or alkali, and this depends on the concrete substituting group of finding on compound as herein described.In the time that compound of the present invention contains relatively acid sense, the required alkali (pure or in suitable inert solvent) of this compounds contact q.s by making neutral form can obtain base addition salt.The example of pharmaceutically acceptable base addition salt comprises sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or class is saloid.In the time that compound of the present invention contains relatively the sense of alkalescence, the required acid (pure or in suitable inert solvent) of this compounds contact q.s by making neutral form can obtain acid salt.The example of pharmaceutically acceptable acid salt comprises derived from those of mineral acid, described mineral acid example hydrochloric acid, Hydrogen bromide, nitric acid, carbonic acid, one hydrogen carbonic acid (monohydrogencarbonic acid), phosphoric acid, one hydrogen phosphoric acid (monohydrogenphosphoric acid), dihydrogen phosphoric acid (dihydrogenphosphoric acid), sulfuric acid, one hydrosulphuric acid (monohydrogensulfuric acid), hydroiodic acid HI or phosphorous acid etc., and derived from relatively nontoxic organic acid salt, described organic acid is as acetic acid, propionic acid, isopropylformic acid, oxalic acid, toxilic acid, propanedioic acid, phenylformic acid, succsinic acid, suberic acid, fumaric acid, amygdalic acid, phthalic acid, Phenylsulfonic acid, tosic acid, citric acid, tartrate, methylsulfonic acid etc.Also comprise amino acid whose salt, described amino acid is as arginine etc., and organic acid salt, and described organic acid is as glucuronic acid or galacturonic acid (galactunoric acid) etc.Some specific compound of the present invention contains alkalescence and acid functional simultaneously, and it makes compound can change into base addition salt or acid salt.
The compound of neutral form can be regenerated by making described salt also separate in a usual manner parent compound with alkali or acid contact.The parent form of compound is in some physical properties---as the solubleness in polar solvent---and upper different from various salt forms, but in other respects, for the object of the invention, the parent form of salt and compound is equal to.
Except salt form, described compound can be prodrug forms.The prodrug of compound is for issuing biochemical variation so that those compounds of compound of the present invention to be provided at physiological condition.In addition, prodrug changes into compound of the present invention by chemistry or biochemical method in environment in vitro.For example, when be placed in contain suitable enzyme or chemical reagent through skin patch holder time, prodrug can slowly change into compound of the present invention.Prodrug is normally useful, because in some cases, it is easier to administration than parent drug.For example, compared with parent drug, it can have more bioavailability by oral administration.With respect to parent drug, prodrug also can have the solubleness in pharmacology composition of improvement.Various prodrug derivants are known in the art, as relied on those of the hydrolytic rupture of prodrug or oxidized activating.The example of prodrug is (but being not limited to) compound of the present invention, and it gives as ester (described " prodrug "), but metabolism is hydrolyzed into carboxylic acid subsequently---active entity.
The form that titled reference compound can be non-solvated and the form of solvation (comprising hydrated form) exist.Conventionally, the form of solvation is equal to non-solvated form, and is intended within the scope of the present invention.Described compound can polycrystalline or the existence of unbodied form.Conventionally, all physical form are equal to for desired use, and are intended within the scope of the invention.
Some titled reference compound has unsymmetrical carbon (optical center) or two key; Racemic modification, diastereomer, geometrical isomer and individual isomer are all intended within the scope of the present invention.
Can take the form of bulk liquid solution or suspension or body powder for the composition of administration.But more commonly, composition exists with unit dosage, to contribute to accurate-metering.Term " unit dosage " refers to and is suitable as unitary dose for human experimenter and other mammiferous physics discrete unit, and constituent parts contains the active substance that can produce as calculated the predetermined amount of required result for the treatment of, and suitable drug excipient.Common unit dosage comprise liquid composition be pre-charged with, the peace measured is cutd open bottle or syringe in advance, or for the pill of solids composition, tablet, capsule, lozenge (losenge) etc.
Various suitable preparations and delivery system (comprising suitable vehicle or carrier) and be known or be significantly to those skilled in the art for the preparation of the method for the composition that can give, and be documented in more detail if Remington:The Science and Practice of Pharmacy is (in the publication of Pharmaceutical Press (2012).For example, in specific embodiments, composition is prepared or is sent in that extend or controlled delivery system, described delivery system for example, as diffusion system (, the implants of storing unit, matrix device, diffusion-controlled and through skin patch) and capsule and stromatolysis system, corrode product, osmotic pump system, ion exchange resin etc.
In specific embodiments, dosage far exceedes the current dosage for parkinsonian defined (200mg), and be preferably 0.5-10,0.5-5,0.5-2.5,1-10,1-5,1-2.5,2-10 or 2-5g/ days, with 0.25,0.5,1,1.5,2 or the unit dosage of 2.5g carry out administration.
For the processing of preventing and/or treating property, compound can give by the whole bag of tricks, includes but not limited to, administered parenterally, local topical administration, oral administration or topical, as passed through aerosol or percutaneous dosing.In addition, according to experienced clinicist's knowledge, the reaction observing to given therapeutical agent of the effect that observe of visual the given therapeutical agent for the treatment of plan to patient and disease and changing.
Should be understood that embodiment described herein and embodiment are only for illustration purpose, and its various amendments or variation are in those skilled in the art's spirit and scope that can expect and that be included in the application and within the scope of the appended claims.All publications, patent, the patent application (comprising its quoted passage) of quoting are herein included in herein by the mode of reference in full for all objects.
Embodiment: the investigation of the antiseptic-germicide skeleton of the discovery of the phentriazine-bis--N-oxide compound to rf and non-replicating mycobacterium tuberculosis target and the activation of optimization biological reducing.Select the compound (that is, needing electronics to carry out enzyme transfer and there is the compound of cytoactive from reductase enzyme) of four class biological reducings activation, to filter out the activity of opposing Mtb H37Rv.Described compound comprises 2-nitroimidazole and 5-nitroimidazole, 2-nitrofuran, quinoxaline-Isosorbide-5-Nitrae-bis--N-oxide compound and BTO (Table A).The compound of these particular types is to select according to the existing report to known anti-microbial activity and according to commercial source availability or National Cancer Institute (National Cancer Institute, NCI) treatment development division (Developmental Therapeutics Program) compound library. 8-10we are according to the MIC value of the Mtb H37Rv bacterial strain in two kinds of sources of compound opposing and restore test (low-oxygen recovery assay, LORA) according to it at hypoxemia 11in (table 1), the MIC value of opposing NRP Mtb is carried out SCREENED COMPOUND.Owing to reporting that H37Rv laboratory strains exists multiple variation 12, primary screen is selected in two independently to carry out with the H37Rv bacterial strain of different sources in laboratory, to confirm actively not only have specificity for given laboratory strains.In addition, we have tested the cytotoxicity of all compound opposing Vero cells, have obtained selectivity index (SI), and it represents with respect to mammal cell line, the relative cytotoxicity of compound to Mtb.
Table A. there is the chemical structure of the biological reducing activating compounds of known anti-microbial activity.
The tuberculosis activity of the different skeletons of table 1. biological reducing activation antimicrobial compounds.
anA=does not obtain; Institute of tuberculosis of University of Illinois, ITR=Chicago.
bthe calculating of carrying out for the Z stereochemistry around two keys.
From the enquiry data demonstration of biological reducing activation skeleton, the BTO of test has the constant activity of opposing active replication type and non-replicating Mtb.The MIC value of opposing active replication type H37Rv bacterial strain is that 0.57-5 μ g/mL and the MIC value in the LORA of NRP model are 0.37-7.3 μ g/mL.All compounds all have some cytotoxicities of the opposing Vero cell of medium SI value.Utilize the result of initial investigation, we have selected the anti-TB activity of the analogue of BTO, particularly 8a to carry out SAR exploitation and have optimized research.
The BTO's that N-replaces is synthetic and active.The 3-amine position that the first new BTO being combined into is included in ring has the different analogues (scheme 1) that replace.Effect and the selectivity how spatial variations in order to be determined at this position and electronic variable affect compound in the replacement of this position.By making primary amine initial substance (table B) react the analogue of synthetic modified side chain with the chloro-phentriazine-1-N-of 3-oxide compound (9).Carry out amine oxide affixture with full trifluoroacetic acid subsequently, described trifluoroacetic acid by adding the preparation of trifluoroacetic anhydride (TFAA) original position in the stirred solution of the compound of excess hydrogen peroxide.Carry out the complete oxidation of 3-4 days by the TFAA that adds extra equivalent, until detected oxidation by TLC.
Scheme 1.N-alkyl 1,2,4-phentriazine-bis--N-oxide compound synthetic.Reagent and condition: (a) RNH 2(2 equivalent), TEA (2 equivalent), DCM, 18h, room temperature, (b) TFAA, H 2o 2, THF, 4 days.
Table B. is for the various kinds of amine unit of phentriazine side chain.
Result shows to have tolerance at non-annularity and the cyclic alkyl of all size of 3-amine position, but do not have modification significantly to increase the SI of compound.(table 2) all compounds all have activity (MIC≤10 μ g/mL); Tool activity be those compounds (8i and 8m) with very hydrophobic side chain.In side chain, include polar ether group (8i) in and cause active significantly reduction, but cytotoxicity also there is suitable reduction.Whole presentation of results of this group are at the ring-type of this position and the more hydrophobic group of non-annularity substituting group tolerance, obtain compounds effective.In this group, do not have modification to improve significantly the SI of compound, described SI is 2.9-8.3.
The tuberculosis activity of the BTO of table 2. side substitution
Cyclosubstituted BTO's is synthetic and active.The mechanism of action of these BTO is considered to, and ring is cytotoxicity radical species by biological reducing activation, and described cytotoxicity radical species causes the DNA damage of the unrepairable of Mtb.In view of this mechanism, the electron reduction current potential of compound should determine Compound Phase for mammalian cell the Cytotoxic selectivity to Mtb.On heterocycle, substituent variation can change significantly electron reduction current potential and affect the selectivity index of compound.
In the time can utilizing, start from the aniline initial substance (11) replacing or the 2-N-methyl-p-nitroaniline (12) replacing for the preparation of the synthetic route of the BTO that encircles modification.(scheme 2) is by aniline acidylate, nitrated with nitrosonitric acid, subsequently by refluxing and go protection in hydrochloric acid, to obtain one group of 2-N-methyl-p-nitroaniline (12). 13subsequently 2-N-methyl-p-nitroaniline in concentrated hydrochloric acid with cyanamide condensation, next use alkaline purification, to form 1,2,4-phentriazine-(1N)-oxide compound ring.The amino phentriazine of 3-changes into 3-hydroxy benzo triazine by react formation diazonium compound intermediate (it is acid hydrolysis) with Sodium Nitrite or nitrite tert-butyl subsequently.This intermediate carries out chlorination with preparation 3-chlorine phentriazine-(1N)-oxide intermediate (14a-g) by refluxing in phosphoryl chloride.Intermediate 14a-g (shows B with the subgroup of primary amine side chain subsequently; Side chain a, b, d, e, f, i and m) reaction.Select side chain a, b, d and f, this is because these low molecular weight side chains obtain compounds effective without increasing unnecessary extra lipophilicity. 14select side chain i with the polar group sampling to this position, select m, this is because it obtains one of compounds effective from the analogue of a series of side substitution.Subsequently, as described in scheme 2, cyclosubstituted this group BTO is oxidized, to obtain compound listed in table 3.
Scheme 2. is cyclosubstituted 1,2,4-phentriazine-bis--N-oxide compound synthetic.Reagent and condition: (a) Ac 2o, TEA, DCM, (b) fHNO 3, Ac 2o, DCM, (c) HCl, refluxes, (d) NCNH 2, dense HCl, (e) NaNO 2, HCl, (f) tBuONO, tBuOH, H 2o, (g) POCl 3, reflux, (h) RNH 2(2 equivalent), TEA (2 equivalent), DCM, 18h, room temperature, (i) TFAA, H 2o 2, THF, 4 days.
Test tuberculosis activity and the cytotoxicity (table 3) of all cyclosubstituted BTO.Cyclosubstituted modification is caused to effect (0.15-5 μ g/mL) and the cytotoxicity (< 2.5-100 μ g/mL) of wide region.The most important thing is, some rings replace the remarkable rising that causes SI value.The general trend of observing is to replace and make the cytotoxicity of mammalian cell reduce to electronics.This trend matches with the hypothesis that the single electron reduction potential (E1/2) that reduces compound can effectively increase SI value. 15the best compound of this series is on benzotriazole ring, to have little alkyl to replace those of (as 5-methyl and 6,7-cyclopentyl replace).These member ring systems produce near the compound of SI 50.
The tuberculosis activity of the cyclosubstituted BTO of table 3.
Two-N-alkyl BTO's is synthetic and active.From the SAR data declaration of side chain and cyclosubstituted compound, around phentriazine ring, increase electron-donating group has improved selectivity index, and the modification of alkylamine side chain has been increased to effect, but remarkably influenced SI not.From these are observed, the conclusion that we draw is to synthesize the new compound on alkylamine side chain with more electron-donating groups.In previous trial, can not synthesize the compound with three replacement amine side chains, this is because final oxidation reaction condition does not produce required two-N-oxide products.In order to prepare these compounds, therefore we developed the oxygenant of some alternative full trifluoroacetic acids, identifies HOFCH 3cN, as the enough strong reagent that carries out required conversion, can not produce significant peroxidation by product. 16,17this variation route allows synthetic a series of two-N-alkyl BTO (scheme 3).For this route, proved to be preferably oxidized 3-chlorine phentriazine intermediate, thereby amine variation step is to carry out the synthetic final step in laboratory by common intermediate 16 and 19.
Scheme 3. 2-N-alkyl 1,2,4-phentriazine-bis--N-oxide compound synthetic.Reagent and condition: (a) F 2(g), H 2o, MeCN, (b) RNH 2(2 equivalent), TEA (2 equivalent), DCM, 1h, room temperature.
In the time carrying out tuberculosis activity and cytotoxicity test, find that new two-N-alkyl BTO has the SI (table 4) of increase.The tuberculosis of compound is active conventionally quite active with the tuberculosis of cyclosubstituted analogue, but the cytotoxicity of opposing Vero cell significantly reduces.From this group compound, we identify the new compound of some SIs > 50.From the secondary amine initiator group (scheme 3) for this group compound, find that little alkyl has the highest activity.Add the polarity or the hydrophilic radical that are intended to for increasing solubleness to cause the active significantly decline of anti-TB.
The tuberculosis activity of table 4. two-N-alkyl BTO
The activity of antagonism property of medicine Mtb bacterial strain.Whether we have also tested to one group of Mtb bacterial strain with single medicine resistance the most effectively and optionally one group of BTO (table 5) of tool, have crossed resistance and keep its effect to multiple Mtb bacterial strain to measure these compounds with existing TB medicine.The data obtained shows, crossed resistance does not occur, and these compounds have effect Mtb drug-fast strain being equated or increase.
Table 5.BTO is to having the MIC of Mtb bacterial strain of single medicine resistance
The antimicrobial spectrum of BTO activity.Consider longer treatment time and complicated drug regimen scheme, antitubercular agent should have narrow anti-microbial activity spectrum.Introduce the anti-microbial activity (SI table 1) of two kinds of BTO---8a and 20q---to different Gram-positives and the gram negative bacterium pathogenic agent of one group.Strikingly, BTO has high selectivity to the activity of mycobacterium, Mtb is had compared with strong activity and M. smegmatics (M.smegmatis.) mycobacterium abscessus (M.abscessus) had weak active, M. smegmatics mycobacterium abscessus has natural resistance to many class microbiotic, and 8a and 20q are all had to resistance. 18
The sudden change potential of BTO.In the antibacterials of any biological reducing activation of exploitation, tend to have the potential of sudden change.Because the active basis of these compounds is to produce intermediate radical species, unwanted sudden change brings risk, as the Bioreductive drugs for many classes of having recorded, comprises BTO Win-59075 (TPZ). 19,20the method of the most convenient of the potential that assessment induced gene damages is to use dull and stereotyped bulk testing method (plate incorporation method), the method is used salmonella typhimurium strain TA98 and TA100, is commonly referred to Ames (Ames) test.Salmonellas test strain has sudden change on histidine operon, this sudden change causes defective lipopolysaccharides (rfa), and be encompassed in vitamin H (Bio) synthetic in and the disappearance of the gene that relates in the reparation (uvrB) that damages at the DNA of uv induction. 21,22these sudden changes make bacterial strain be increased and increase its susceptibility to these molecule mutagenic effects to permitting polymolecular perviousness.The postulated mechanism of considering BTO compound is optionally to form with respect to mammalian cell and have Cytotoxic radical species in bacterium, and microbiologic mutation test is not desirable, but in its activity of benchmark test for this test.In the case of there is and do not exist the mouse liver metabolism activation system of the Aroclor1254 induction that contains 10%S9 (MA), in Ames test, assessment has two kinds of BTO, 15fa and the 20q of good efficacy and selectional feature.The condition of single plate count and mean value thereof and standard deviation and background lawn (background lawn) is shown in SI table 2.In the situation that existing and do not have metabolism activation, the increase of the recovery mutation colony number of two kinds of bacterial strains of TA98 and TA100 is relevant to the dosage of 15fa and 20q.In order to assess it at the sudden change potential without in microorganism system, also at mouse lymphoma cell tk +/-→ tk +/-in transgenation (MOLY) test, assess 15fa and 20q, this test is a kind of conventional genetic toxicology test (table 6) of the potential that suddenlys change in mammalian cell for assessment of compound. 23,24in the situation that existing or not having metabolism activation (S9), the 15fa under the non-cell toxicity dosage level of 5,10,25 and 50 μ g/mL demonstrates the mutation frequency MF of remarkable rising.For 20q, in the situation that existing or not having S9, under the non-cell toxicity dosage level that is up to 100 μ g/mL, expose 4 hours, mutation frequency does not increase, in the situation that not there is not S9, under the non-cell toxicity dosage level that is up to 10 μ g/mL, expose 24 hours, mutation frequency does not increase.Generally speaking, exist and there is not S9 in the situation that, 20q is to the mutation frequency response that is negative.
The sudden change potential of BTO in table 6.MOLY test.(S9=metabolism of rat activation; Positive=significantly induced mutation frequency; Feminine gender=non-remarkable induced mutation frequency; NT=does not test)
ainduced mutation frequency, but for average solvent control group a little higher than 40x10 -6net value, biologically thinks and is negative.
Plysiochemical characteristic and mouse pharmacokinetics.In order to assess the potential of BTO as the therapeutical agent of oral bioavailability, plysiochemical characteristic and the ADME characteristic of 8a and 20q described.In the solubleness of these two kinds of compounds adopts fask oscillating method be 7.4 at pH 0.9% normal saline solution, measure three times.8a and 20q have respectively the acceptable solubleness of 1.35mg/mL and 0.28mg/mL.
In order to verify that BTO is as the new leading series that is used for the treatment of TB, we have also assessed its general in mouse after oral administration and have exposed.In order to describe the pharmacokinetics of BTO, give 8a and the 20q (table 7) of single dose (100mg/kg) to mouse.For in fixed dosage composition tablet with compatible mutually when last line TB medicine, our target drug candidate feature is oral administration therapeutical agent, its maximal dose needing is for once a day, preferred dose is one week 1-3 time. 25in order to realize this feature, in our development project, taking elimination transformation period of the mouse of 4-12 hour as target.The elimination transformation period of every kind of compound is all shorter than our dreamboat value, but T maxvalue shows that it is rapidly absorbed (Fig. 1) after oral administration.According to C maxwith AUC value, two kinds of compounds all produce good exposure, and this shows that these compounds promise to be antitubercular agent.
The pharmacokinetic parameter of table 7. to 8a after female mice oral administration and 20q.
Compound Dosage t 1/2 T Maximum C Maximum AUC Finally AUC Infinite
? (mg/kg) (hr) (hr) (ng/ml) (hr·ng/ml) (hr·ng/ml)
8a 100 1.5 0.08 9540 6578 6937
20q 100 1.9 0.25 9767 11219 11248
Our SAR shows, hydrophobic side chain produces more effective tuberculosis compound conventionally, and this expects, because compound must be by hydrophobic its activity of mycobacterium cell walls competence exertion.Increasing hydrophobicity and also seem to have increased the cytotoxicity to mammalian cell, may be by increasing cell permeability.The modification of the replacement on BTO ring causes the optionally substantial variation of compound.Conventionally, the introducing of electron-withdrawing group (as halogen) causes compound to more effectively (MIC is 0.15-0.31 μ g/mL) of Mtb, but these compounds also have more cytotoxicity to Vero cell.The replacement to electronics on ring tends to slightly reduce the effect (MIC is 0.31-1.2 μ g/mL) to Mtb, but has substantially reduced toxicity, produces the compound of the selectivity feature with comprehensive improvement.For these two kinds contrary SAR trend of balance, we have selected to have the compound (15f) of fused tricyclic system, and it has best balanced effect and feature optionally.
The development of the oxidation chemistry of preparation two-N-alkyl BTO compound makes us can prepare the compound that anti-Mtb effect improves 2-4 times, keeps or the toxicity of reduction to Vero cell simultaneously.Except having the compound of the more hydrophobic side chain that contains morpholine (17p), sulfone (20v) or sulphonamide (20x), all compounds of preparing in this series have extraordinary effect (MIC is 0.31-0.62 μ g/mL); All these compounds demonstrate significantly reduced tuberculosis activity (MIC is 2.5-5 μ g/mL).In a word, SAR trend explanation, optimum compound should contain electron rich member ring systems with hydrophobic side chain, and (E1/2 value is lower; LUMO energy is higher), to obtain having maximum effect and compound optionally.
The plysiochemical characteristic of 8a and 20q and the analysis of mouse pharmacokinetic data show, this class has good class pharmaceutical properties.The solubleness of described compound is conventionally very good, and this is likely because the polarity essence of the N-oxide groups on phentriazine ring.It is active that 20q does not have sudden change under the concentration that is up to 100 μ g/mL.
The general procedure of amination/oxidation.Method A.At room temperature, primary amine (2.2mmol) initial substance is joined in the stirred solution that is dissolved in 9 (200mg, 1.1mmol) in the there is triethylamine DCM (10mL) of (5.5mmol).By solution stirring 24-48 hour, all consume until monitor 9 all initial substances by TLC (5%MeOH/DCM moving phase).Body amine side chain need to be heated to 45 DEG C to transform completely.Whole reaction vapourisation under reduced pressure, to obtain crude product 10a-n, it is directly through oxidizing condition processing.By thick list-N-oxide dissolution at THF (8mL) and 50%H 2o 2(4mL) in mixture, use subsequently TFAA (3.3mmol) slowly to process, at room temperature stir simultaneously.Monitor oxidation by TLC, and every TFAA (3.3mmol) that adds extra equivalent for 12 hours, until reacted > 90% or no longer formed extra product.When reaction finishes, reaction is with the saturated sodium bicarbonate cancellation of 10mL and with the chloroform extraction of 10mL three times.The organic layer of merging is evaporated and carries out purifying by semi-preparative reversed-phase HPLC.
The general procedure of the amino phentriazine-1-oxide compound of 3-.Method B.In the three-necked bottle of 1L that is equipped with mechanical stirring and reflux exchanger, put into 2-N-methyl-p-nitroaniline (12a-h, 171mmol), cyanamide (1.37mol, 8 equivalents) and Et 2o (30mL).Mixture is heated 1 hour at 100 DEG C, be cooled to subsequently 50-55 DEG C.Use dropping funnel to drip dense HCl (72mL), simultaneously mechanical stirring (noting: strong exothermal reaction).Add when finishing and not having gas further to overflow, mixture is heated 3 hours at 110 DEG C.Subsequently reaction is cooled to 50 DEG C and also slowly adds NaOH solution (7.5M, 160mL).Reheated 110 DEG C with oil bath subsequently, continued 3 hours, cooling and pour the H of 3L into 2in O.Filter gained solid, use H 2o and Et 2o washing, dry under vacuum subsequently.If needed, product is by purified by flash chromatography, and described flash chromatography uses silica gel, the CH of the MeOH that eluent gradient is 0-10% 2cl 2solution.
The general procedure of 3-chlorine phentriazine-1-oxide compound.Method C.At room temperature, by NaNO 2(1.5g, 21mmol) joins in trifluoroacetic acid (15mL) stirred solution of the amino phentriazine-1-oxide compound of 3-(13a-e, 7mmol) with aliquot.After 1 hour, add the H of 50mL 2o, and filter out solid, and dry under vacuum.The POCl at 20mL by solid suspension subsequently 3in and reflux 3 hours.Cooling solution, removes most of excessive POCl by distillation 3, and be dissolved in subsequently CH 2cl 2(40mL) in.Use subsequently H 2o and this solution of salt water washing, and evaporating solvent.Thick residue is by purified by flash chromatography, and described flash chromatography uses the CH of the EtOAc that silica gel and gradient are 0-10% 2cl 2solution.
The general procedure of 3-chlorine phentriazine-1-oxide compound.Method D.To tBuOH (50mL) and the H of the amino phentriazine-1-oxide compound of 3-(13f-g, 9mmol) 2in the stirred solution of O (5mL), slowly add tBuONO (90%, 15mL) and H 2sO 4(1mL).Be heated to 60 DEG C spend the night after, mixture is joined in the ice of 100g, filter gained solid, and dry under vacuum.The POCl at 20mL by solid suspension subsequently 3in, and reflux 3 hours.Cooling solution, removes most of excessive POCl by distillation 3, and be dissolved in subsequently CH 2cl 2(40mL) in.Use subsequently H 2o and this solution of salt water washing, and evaporating solvent.Thick residue is by purified by flash chromatography, and described flash chromatography uses the CH that silica gel and gradient are 0-10%EtOAc 2cl 2solution.
HOFCH 3the general procedure of CN oxidation.Method E.(note: F 2and HOFCH 3cN is extremely strong oxygenant and corrosives) at-15 DEG C, by 10% F that contains nitrogen 2slowly bubbling is to containing 10mL H 2the CH of the 100mL of O 3in CN, continue 1 hour.To being dissolved in CH 2cl 2(80mL) the disposable oxygenant that adds gained in cooling (0 DEG C) solution of the phentriazine-1-N-oxide compound (12.6mmol) in.Mixture stirs 10 minutes, uses subsequently saturated NaHCO 3(40mL) cancellation reaction.Two-phase mixture is used H subsequently 2o (40mL) dilutes and uses CHCl 3(80mL) extraction.Organic layer is used H subsequently 2o, salt water washing, and use MgSO 4dry.Evaporating solvent, product is by purified by flash chromatography, and described flash chromatography uses the CH of the EtOAc that silica gel and gradient are 0-10% 2cl 2solution.The yield separating is 35-40%, and surplus materials is recovered as to initial substance.
The general procedure of the amination of 3-chlorine phentriazine-Isosorbide-5-Nitrae-bis--N-oxide compound.Method F.At 0 DEG C, in the stirred solution of the glycol dimethyl ether of 3-chlorine phentriazine-Isosorbide-5-Nitrae-bis--N-oxide compound (2mmol), add amine (2.2mmol) and triethylamine (2.4mmol).Mixture is warming up to room temperature and stirs 16 hours.After reaction, evaporating solvent, and crude product passes through purified by flash chromatography, the CH that described flash chromatography use silica gel and gradient are 0-10%MeOH 2cl 2solution.Common yield is 80-90%.
3-(ethylamino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8a).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400?MHz)：δ8.28(br-s，1H)，8.18(d，J＝8.4Hz，1H)，8.10(d，J＝8.5Hz，1H)，7.90(t，J＝7.5Hz，1H)，7.55(t，J＝8.6Hz，1H)，3.43(m，2H)，1.18(t，J＝7.0Hz，3H).MS(ESI+)：m/z207.0((M+H) +)。
3-(cyclopropylamino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8b).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.41(d，J＝2.4Hz，1H)，8.20(dd，J＝8.8，0.8Hz，1H)，8.10(dd，J＝8.8，0.8Hz，1H)，7.92(m，1H)，7.56(m，1H)，2.76(m，1H)，0.74(m，4H)。MS(ESI+)：m/z219.0((M+H) +)。
3-(tertiary butyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8c).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.31(d，J＝8.8Hz，1H)，8.26(d，J＝8.8Hz，1H)，7.83(t，J＝7.0Hz，1H)，7.45(t，J＝7.0Hz，1H)，7.19(s，1H)，1.55(s，9H)。MS(ESI+)：m/z235.0((M+H) +)。
3-(cyclobutyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8d).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.44(d，J＝8.0Hz，1H)，8.17(dd，J＝8.8，0.8Hz，1H)，8.10(dd，J＝8.8，0.8Hz，1H)，7.91(m，1H)，7.54(m，1H)，4.33(m，1H)，2.24(m，4H)，1.67(m，2H)。MS(ESI+)：m/z233.0((M+H)+)。
3-(cyclopentyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8e).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.18(dt，J＝8.8，0.8Hz，1H)，8.10-8.06(m，2H)，7.92(m，1H)，7.54(m，1H)，4.18(m，1H)，1.92(m，2H)，1.70(m，4H)，1.56(m，2H)。MS(ESI+)：m/z247.0((M+H)+)。
3-(cyclohexyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8f).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.16(dd，J＝8.8，0.8Hz，1H)，8.08(dt，J＝8.8，0.8Hz，1H)，7.95(d，J＝9.2Hz，1H)，7.89(m，1H)，7.52(m，1H)，3.71(m，1H)，1.85(m，2H)，1.72(m，2H)，1.58(m，1H)，1.51-1.42(m，2H)，1.35-1.26(m，2H)，1.20-1.09(m，1H)。MS(ESI+)：m/z261.0((M+H)+)。
3-(suberyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8g).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.18(dt，J＝8.8，0.8Hz，1H)，8.09(dt，J＝8.8，0.8Hz，1H)，7.96(d，J＝8.4Hz，1H)，7.91(m，1H)，7.53(m，1H)，3.92(m，1H)，1.89(m，2H)，1.76-1.42(m，10H).MS(ESI+)：m/z275.0((M+H)+)。
3-(penta-2-base amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8h).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.16(dt，J＝8.8，0.4Hz，1H)，8.07(dt，J＝8.8，0.4Hz，1H)，8.00(d，J＝9.2Hz，1H)，7.90(m，1H)，7.52(m，1H)，3.97(m，1H)，1.65(m，1H)，1.49(m，1H)，1.30(m，2H)，1.20(d，J＝6.4Hz，3H)，0.83(t，J＝7.2Hz，3H)。MS(ESI+)：m/z249.0((M+H)+)。
3-((1-methoxyl group fourth-2-yl) amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8i).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.18(dt，J＝8.8，0.8Hz，1H)，8.09(dt，J＝8.8，0.8Hz，1H)，7.92(m，2H)，7.54(m，1H)，3.50(dd，J＝10，6.4Hz，1H)，3.40(dd，J＝10，5.2Hz，2H)，3.23(s，3H)，1.62(m，2H)，0.87(m，3H)。MS(ESI+)：m/z265.0((M+H)+)。
3-((4-(tertiary butyl) cyclohexyl) amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8j).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.17(td，J＝8.4，0.8Hz，1H)，8.08(m，1H)，7.97(d，J＝9.2Hz，1H)，7.91(m，1H)，7.57-7.50(m，1H)，4.06(m，1H)，1.94(m，2H)，1.75(m，1H)，1.58(m，2H)，1.14(m，4H)，0.83(d，J＝3.2Hz，9H)。MS(ESI+)：m/z317.1((M+H)+)。
(R)-3-((1-cyclohexyl ethyl) amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8k).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.16(dd，J＝8.8，0.8Hz，1H)，8.08(dd，J＝8.8，0.8Hz，1H)，7.89(m，2H)，7.51(m，1H)，3.75(m，1H)，1.72-1.55(m，6H)，1.18(d，J＝6.8Hz，3H)，1.12(m，3H)，0.92(m，2H)。MS(ESI+)：m/z289.0((M+H)+)。
3-((5-methyl oneself-2-yl) amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8l).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.16(dd，J＝8.8，0.8Hz，1H)，8.09(dd，J＝8.4，0.4Hz，1H)，7.94(d，J＝9.6Hz，1H)，7.89(m，1H)，7.52(m，1H)，3.91(m，1H)，1.66(m，1H)，1.50(m，2H)，1.20(d，J＝6.8Hz，3H)，1.15(m，2H)，0.83(dd，J＝6.4，2.0Hz，6H)。MS(ESI+)：m/z277.0((M+H)+)。
3-((1R, 2R, 4S)-dicyclo [2.2.1] heptan-2-base amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8m).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.18(dt，J＝8.8，0.4Hz，1H)，8.09(dd，J＝8.8，0.8Hz，1H)，7.90(m，1H)，7.73(d，J＝6.8Hz，1H)，7.53(m，1H)，3.65(m，1H)，2.26(m，2H)，1.69(m，2H)，1.50(m，3H)，1.14(m，3H)。MS(ESI+)：m/z273.0((M+H)+)。
3-((1R, 3S, 5r, 7r)-diamantane-2-base amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (8n).Synthetic by 9 by method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.18(dd，J＝8.8，0.8Hz，1H)，8.11(dd，J＝8.8，1.2Hz，1H)，7.93(m，1H)，7.55(m，1H)，7.39(d，J＝8.4Hz，1H)，4.00(m，1H)，2.04(m，2H)，1.88(m，8H)，1.72(br?s，2H)，1.61(m，2H)。MS(ESI+)：m/z313.1((M+H)+)。
3-(ethylamino)-7-fluoro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15aa).Synthetic by the fluoro-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(CDCl 3，400MHz)：δ8.26(dd，J＝4.9，9.6；1H)，7.92(dd，J＝1.7，8.0；1H)，7.57(m，1H)，6.94(br-t，1H)，3.57(p，J＝7.3，2H)，1.29(t，J＝7.3，3H)。MS(ESI+)：m/z224.9((M+H)+)。
3-(cyclopropylamino)-7-fluoro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ab).Synthetic by the fluoro-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.37(br?s，1H)，8.15(dd，J＝9.6，5.2Hz，1H)，7.97(dd，J＝8.4，2.8Hz，1H)，7.84(m，1H)，2.72(m，1H)，0.72(m，4H)。MS(ESI+)：m/z236.9((M+H)+)。
3-(cyclobutyl amino)-7-fluoro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ad).Synthetic by the fluoro-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.43(d，J＝8.0Hz，1H)，8.15(dd，J＝9.6，5.2Hz，1H)，7.93(dd，J＝8.8，2.8Hz，1H)，7.83(m，1H)，4.30(m，1H)，2.23(m，4H)，1.67(m，2H)。MS(ESI+)：m/z250.9((M+H)+)。
3-(cyclopentyl amino)-7-fluoro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ae).Synthetic by the fluoro-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.14(dd，J＝9.6，5.2Hz，1H)，7.97(m，1H)，7.94(dd，J＝8.4，2.8Hz，1H)，7.83(m，1H)，4.13(m，1H)，1.91(m，2H)，1.67(m，4H)，1.55(m，2H)。MS(ESI+)：m/z265.0((M+H)+)。
The fluoro-3-of 7-((1-methoxyl group fourth-2-yl) amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ag).Synthetic by the fluoro-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.16(dd，J＝9.6，5.2Hz，1H)，7.95(dd，J＝9.2，2.8Hz，1H)，7.85(m，2H)，3.95(m，1H)，3.49(dd，J＝10，6.4Hz，1H)，3.39(dd，J＝9.6，5.2Hz，1H)，3.23(s，3H)，1.61(m，2H)，0.86(t，J＝7.2Hz，3H)。MS(ESI+)：m/z283.0((M+H)+)。
3-(cyclohexyl amino)-7-fluoro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15aj).Synthetic by the fluoro-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.14(dd，J＝9.2，5.2Hz，1H)，7.95(dd，J＝8.8，2.8Hz，1H)，7.91(m，1H)，7.83(m，1H)，3.69(m，1H)，1.84(m，2H)，1.71(m，2H)，1.58(m，1H)，1.51-1.41(m，2H)，1.35-1.25(m，2H)，1.12(m，1H)。MS(ESI+)：m/z279.0((M+H)+).
3-((1R, 2R, 4S)-dicyclo [2.2.1] heptan-2-base amino)-7-fluoro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15an).Synthetic by the fluoro-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid.1H-NMR(DMSO-d 6，400MHz)：δ8.15(dd，J＝9.6，4.8Hz，1H)，7.95(dd，J＝9.2，2.8Hz，1H)，7.84(m，1H)，7.74(d，J＝7.2Hz，1H)，3.61(m，1H)，2.28(d，J＝4.0Hz，1H)，2.22(brs，1H)，1.69(m，2H)，1.56(m，1H)，1.45(m，2H)，1.12(m，3H)。MS(ESI+)：m/z291.0((M+H)+).
The chloro-3-of 7-(ethylamino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ba).Synthesized by 4-chloro-2-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.41(t，J＝6.0Hz，1H)，8.19(d，J＝2.4Hz，1H)，8.10(d，J＝9.2Hz，1H)，7.91(dd，J＝9.2，2.4Hz，1H)，3.41(m，2H)，1.17(t，J＝7.2Hz，3H)。MS(ESI+)：m/z241.0((M+H)+)。
The chloro-3-of 7-(cyclopropylamino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15bb).Synthesized by 4-chloro-2-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.47(br?s，1H)，8.19(d，J＝2.0Hz，1H)，8.09(d，J＝9.2Hz，1H)，7.90(dd，J＝9.2，2.0Hz，1H)，2.73(m，1H)，0.77-0.67(m，4H)。MS(ESI+)：m/z253.0((M+H)+).
The chloro-3-of 7-(cyclobutyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15bd).Synthesized by 4-chloro-2-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.55(d，J＝7.6Hz，1H)，8.16(d，J＝1.6Hz，1H)，8.09(d，J＝9.2Hz，1H)，7.89(dd，J＝9.2，2.0Hz，1H)，4.31(m，1H)，2.22(m，4H)，1.66(m，2H)。MS(ESI+)：m/z267.0((M+H)+)。
The chloro-3-of 7-((1-methoxyl group fourth-2-yl) amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15bg).Synthesized by 4-chloro-2-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.18(d，J＝2.4Hz，1H)，8.10(d，J＝9.2Hz，1H)，7.96(d，J＝8.8Hz，1H)，7.90(dd，J＝9.2，2.4Hz，1H)，3.97(m，1H)，3.49(dd，J＝9.6，6.4Hz，1H)，3.39(m，1H)，3.23(m，3H)，1.61(m，2H)，0.86(t，J＝7.2Hz，3H)。MS(ESI+)：m/z299.0((M+H)+)。
The chloro-3-of 7-(cyclohexyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15bj).Synthesized by 4-chloro-2-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.17(d，J＝2.4Hz，1H)，8.08(d，J＝9.2Hz，1H)，8.02(d，J＝8.8Hz，1H)，7.89(dd，J＝9.2，2.4Hz，1H)，3.70(m，1H)，1.82(m，2H)，1.70(m，2H)，1.57(m，1H)，1.46(m，2H)，1.31(m，2H)，1.12(m，1H)。MS(ESI+)：m/z295.1((M+H)+)。
3-((1R, 2R, 4S)-dicyclo [2.2.1] heptan-2-base amino)-7-chloro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15bn).Synthesized by 4-chloro-2-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.18(dd，J＝2.4，0.4Hz，1H)，8.08(dd，J＝9.6，0.8Hz，1H)，7.90(dd，J＝9.2，2.4Hz，1H)，7.85(m，1H)，3.62(m，1H)，2.28(d，J＝4.0Hz，1H)，2.22(brs，1H)，1.69(m，2H)，1.56(m，1H)，1.47(m，2H)，1.14(m，3H)。MS(ESI+)：m/z307.1((M+H)+)。
The bromo-3-of 7-(ethylamino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ca).Synthetic by the bromo-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.41(t，J＝6.4Hz，1H)，8.32(m，1H)，8.01(m，2H)，3.40(m，2H)，1.17(t，J＝7.2Hz，3H)。MS(ESI+)：m/z286.9((M+H)+)。
The bromo-3-of 7-(cyclobutyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15cd).Synthetic by the bromo-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.58(m，1H)，8.31(m，1H)，8.02(m，2H)，4.32(m，1H)，2.32(m，4H)，1.67(m，2H)。MS(ESI+)：m/z312.9((M+H)+).
The bromo-3-of 7-(cyclopentyl amino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ce).Synthetic by the bromo-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.32(t，J＝1.2Hz，1H)，8.14(d，J＝8.0Hz，1H)，8.01(m，2H)，4.16(m，1H)，1.92(m，2H)，1.70(m，4H)，1.56(m，2H)。MS(ESI+)：m/z326.9((M+H)+)。
3-((1R, 2R, 4S)-dicyclo [2.2.1] heptan-2-base amino)-7-bromo benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15cn).Synthetic by the bromo-2-N-methyl-p-nitroaniline of 4-with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.33(t，J＝1.2Hz，1H)，8.02(m，2H)，7.89(m，1H)，3.62(m，1H)，2.30(m，1H)，2.23(m，1H)，1.72(m，2H)，1.59-1.40(m，3H)，1.23-1.01(m，3H)。MS(ESI+)：m/z352.9((M+H)+)。
3-(ethylamino)-7-methoxyl group benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15da).Synthetic by 4-methoxyl group-2-N-methyl-p-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.08(t，J＝6.0Hz，1H)，8.04(d，J＝9.6Hz，1H)，7.57(dd，J＝9.6，2.8Hz，1H)，7.46(d，J＝2.4Hz，1H)，3.90(s，3H)，3.38(m，2H)，1.17(t，J＝7.2Hz，3H)。MS(ESI+)：m/z237.0((M+H)+)。
3-(cyclobutyl amino)-7-methoxyl group benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15dd).Synthetic by 4-methoxyl group-2-N-methyl-p-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.25(d，J＝8.0Hz，1H)，8.05(d，J＝9.6Hz，1H)，7.57(dd，J＝9.2，2.8Hz，1H)，7.45(d，J＝2.4Hz，1H)，4.30(m，1H)，3.90(s，3H)，2.22(m，4H)，1.67(m，2H)。MS(ESI+)：m/z263.0((M+H)+).
3-((1R, 2R, 4S)-dicyclo [2.2.1] heptan-2-base amino)-7-methoxyl group benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15dn).Synthetic by 4-methoxyl group-2-N-methyl-p-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.04(d，J＝9.6Hz，1H)，7.56(m，2H)，7.46(d，J＝2.4Hz，1H)，3.90(s，3H)，3.61(m，1H)，2.30(d，J＝4.0Hz，1H)，2.23(br?s，1H)，1.69(m，2H)，1.57-1.35(m，3H)，1.23-1.09(m，3H)。MS(ESI+)：m/z303.0((M+H)+)。
Ethyl-(5-methyl isophthalic acid, 4-dioxy base-benzo [1,2,4] triazine-3-yl)-amine (15ea).Synthetic by 6-methyl-2-N-methyl-p-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.34(t，J＝6.4Hz，1H)，8.05(dq，J＝8.8，0.8Hz，1H)，7.60(m，1H)，7.38(m，1H)，3.40(p，J＝6.8Hz，2H)，2.95(s，3H)，1.18(t，J＝7.2Hz，3H)。MS(ESI+)：m/z221.0((M+H)+)。
3-(cyclobutyl amino)-5-methyl benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ed).Synthetic by 6-methyl-2-N-methyl-p-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.43(d，J＝8.0Hz，1H)，8.04(d，J＝8.8Hz，1H)，7.59(dt，J＝7.2，1.2Hz，1H)，7.38(m，1H)，4.31(m，1H)，2.96(s，3H)，2.27-2.17(m，4H)，1.68(m，2H)。MS(ESI+)：m/z247.0((M+H)+)。
3-(cyclopentyl amino)-5-methyl benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ee).Synthetic by 6-methyl-2-N-methyl-p-nitroaniline with method C and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.05(m，1H)，7.95(d，J＝7.6Hz，1H)，7.59(dt，J＝7.2，1.2Hz，1H)，7.38(dd，J＝8.8，7.2Hz，1H)，4.15(m，1H)，2.96(s，3H)，1.94(m，2H)，1.68(m，4H)，1.57(m，2H)。MS(ESI+)：m/z261.0((M+H)+)。
3-(ethylamino)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15fa).With method D and method A, by 6-nitro-2,3-dihydro-1H-indenes-5-amine is synthetic.Red solid.1H-NMR(CDCl 3，400MHz)：δ8.11(s，1H)，8.07(s，1H)，6.94(br-t，1H)，3.59(p，J＝7.2，2H)，3.09(t，J＝6.7，2H)，3.03(t，J＝7.7，2H)，2.20(p，J＝7.5，2H)，1.33(t，J＝7.3，3H)。MS(ESI+)：m/z247.0((M+H)+)。
Cyclobutyl-(5,8-dioxy base-2,3-dihydro-1H-5,6,8-, tri-azepine-ring penta [b] naphthalene-7-yls)-amine (15fd).With method D and method A, by 6-nitro-2,3-dihydro-1H-indenes-5-amine is synthetic.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.32(d，J＝8.0Hz，1H)，7.99(brs，1H)，7.95(br?s，1H)，4.32(m，1H)，3.07-2.97(m，4H)，2.22(m，4H)，2.07(m，2H)，1.67(m，2H)。MS(ESI+)：m/z273.0((M+H)+)。
3-((1R, 2R, 4S)-dicyclo [2.2.1] heptan-2-base amino)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15fn).With method D and method A, by 6-nitro-2,3-dihydro-1H-indenes-5-amine is synthetic.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.00(br?s，1H)，7.94(br?s，1H)，7.63(d，J＝6.8Hz，1H)，3.63(m，1H)，3.07-2.97(m，4H)，2.30(d，J＝4.0Hz，1H)，2.24(br?s，1H)，2.07(m，2H)，1.70(m，2H)，1.57(m，1H)，1.49(m，2H)，1.15(m，3H)。MS(ESI+)：m/z313.0((M+H)+)。
3-(ethylamino)-6-methoxyl group-7-methyl benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15ga).Synthetic by 5-methoxyl group-4-methyl-2-N-methyl-p-nitroaniline with method D and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.17(t，J＝6.0Hz，1H)，7.99(d，J＝1.2Hz，1H)，7.33(s，1H)，4.00(s，3H)，3.42-3.36(m，2H)，2.26(d，J＝0.8Hz，3H)，1.17(t，J＝7.2Hz，3H)。MS(ESI+)：m/z251.0((M+H)+)。
3-(cyclobutyl amino)-6-methoxyl group-7-methyl benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15gd).Synthetic by 5-methoxyl group-4-methyl-2-N-methyl-p-nitroaniline with method D and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.31(d，J＝8.4Hz，1H)，7.98(m，1H)，7.33(s，1H)，4.31(m，1H)，4.00(s，3H)，2.26(d，J＝0.8Hz，3H)，2.22(m，4H)，1.67(m，2H)。MS(ESI+)：m/z277.0((M+H)+)。
3-((1R, 2R, 4S)-dicyclo [2.2.1] heptan-2-base amino)-6-methoxyl group-7-methyl benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (15gn).Synthetic by 5-methoxyl group-4-methyl-2-N-methyl-p-nitroaniline with method D and method A.Red solid. 1H-NMR(DMSO-d 6，400MHz)：δ8.01(d，J＝1.2Hz，1H)，7.60(d，J＝6.8Hz，1H)，7.32(s，1H)，4.00(s，3H)，3.63(m，1H)，2.33(m，1H)，2.27(s，3H)，2.24(br?s，1H)，1.69(m，2H)，1.49(m，3H)，1.16(m，3H)。MS(ESI+)：m/z317.0((M+H)+)。
3-chloro benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (16).Synthetic by 9 by method E.Yellow solid. 1H-NMR(CDCl 3，300MHz)：δ8.55(d，J＝8.5，1H)，8.48(d，J＝8.6，1H)，8.07(t，J＝8.5Hz，1H)，7.90(t，J＝8.6Hz，1H)。MS(ESI+)：m/z198.0((M+H)+)。
3-is chloro-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (19).Synthetic by 18 by method E.Yellow solid. 1H-NMR(CDCl 3，300MHz)：δ8.34(s，1H)，8.26(s，1H)，3.17(q，J＝7.8Hz，4H)，2.27(quin，J＝7.8，2H)。MS(ESI+)：m/z238.0((M+H)+)。
3-(diethylamino) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (17o).Synthetic by 16 with method F.Red solid. 1H-NMR(CDCl 3，300MHz)：δ8.36(d，J＝7.4Hz，1H)，8.31(d，J＝7.4Hz，1H)，7.85(t，J＝7.4Hz，1H)，7.54(t，J＝7.4Hz，1H)，3.81(q，J＝6.9Hz，4H)，1.32(t，J＝7.0Hz，6H)。(ESI+)：m/z235.0((M+H)+)。
3-morpholino benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (17p).Synthetic by 16 with method F.Red solid. 1H-NMR(CDCl 3，300MHz)：δ8.39(d，J＝8.7Hz，1H)，8.35(d，J＝9.6Hz，1H)，7.90(t，J＝7.0Hz，1H)，7.65(t，J＝7.0，1H)，3.91(m，4H)，3.87(m，4H)。(ESI+)：m/z249.1((M+H)+)。
3-(pyrrolidin-1-yl) benzo [e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (17q).Synthetic by 16 with method F.Red solid. 1H-NMR(CDCl 3，300MHz)：δ8.31(d，J＝5.8Hz，2H)，7.84(t，J＝5.8，1H)，7.49(t，J＝5.8Hz，1H)，4.04(m，4H)，1.98(m，4H)。(ESI+)：m/z233.0((M+H)+)。
3-(pyrrolidin-1-yl)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20q).Synthetic by 19 with method F.Red solid. 1H-NMR(CDCl 3，300MHz)：δ8.11(s，1H)，8.10(s，1H)，4.03(t，J＝6.6，4H)，3.07(t，J＝7.3Hz，2H)，3.04(t，J＝7.5Hz，2H)，2.19(quin，J＝7.4Hz，2H)，1.98(m，4H)。MS(ESI+)：m/z273.0((M+H)+)。
3-(piperidin-1-yl)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20r).Synthetic by 19 with method F.Red solid. 1h-NMR (CDCl 3, 300MHz): δ 8.15 (s, 1H), 8.11 (s, 1H), 3.76 (m, 4H), 3.05 (m, 4H), 2.18 (quartet, J=7.5Hz, 2H), 1.69 (m, 6H).MS(ESI+)：m/z287.0((M+H)+)。
3-(dimethylamino)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20s).Synthetic by 19 with method F.Red solid.1H-NMR (CDCl 3, 300MHz): δ 8.15 (s, 1H), 8.10 (s, 1H), 3.31 (s, 6H), 3.05 (quartet, J=6.9Hz, 4H), 2.15 (quartet, J=7.3Hz, 2H).MS(ESI+)：m/z247.0((M+H)+)。
3-(benzyl (methyl) amino)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20t).Synthetic by 19 with method F.Red solid. 1h-NMR (CDCl 3, 300MHz): δ 8.24 (s, 1H), 8.16 (s, 1H), 7.25-7.35 (m, 5H), 5.1 (s, 2H), 3.17 (s, 3H), 3.12 (quartets, J=7.6Hz, 4H), 2.21 (quartets, J=7.6Hz, 2H).MS(ESI+)：m/z323.0((M+H)+)。
3-(methyl (styroyl) amino)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20u).Synthetic by 19 with method F.Red solid. 1h-NMR (CDCl 3, 300MHz): δ 8.09 (s, 1H), 8.05 (s, 1H), 7.21 (d, J=7.4Hz, 2H), 7.11 (t, J=5.2Hz, 2H), 7.00 (t, J=7.4Hz, 1H), 4.16 (t, J=7.6Hz, 2H), 3.24 (s, 3H), 2.97-3.11 (m, 6H), 2.18 (quartet, J=7.6Hz, 2H).MS(ESI+)：m/z337.0((M+H)+)。
3-(thiomorpholine is for sulfone-1-yl)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20v).Synthetic by 19 with method F.Red solid. 1h-NMR (CDCl 3, 300MHz): δ 8.17 (s, 1H), 8.14 (s, 1H), 4.35 (m, 2H), 3.26 (m, 2H), 3.12 (m, 2H), 2.23 (quartet, J=7.4Hz, 2H).MS(ESI+)：m/z337.0((M+H)+)。
3-(pipecoline-1-yl)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20w).Synthetic by 19 with method F.Red solid. 1h-NMR (CDCl 3, 300MHz): δ 8.16 (s, 1H), 8.12 (s, 1H), 4.18 (t, J=7.0Hz, 1H), 4.18 (d, J=13.2Hz, 1H), 3.32 (m, 1H), 3.03 (m, 4H), 2.16 (quartet, J=7.3Hz, 2H), 1.92 (m, 1H), 1.62 (m, 6H), 1.33 (d, J=7.0Hz, 3H).MS(ESI+)：m/z301.0((M+H)+)。
3-(4-(methyl sulphonyl) piperazine-1-yl)-7,8-dihydro-6H-indeno [5,6-e] [1,2,4] triazine Isosorbide-5-Nitrae-dioxide (20x).Synthetic by 19 with method F.Red solid. 1h-NMR (CDCl 3, 300MHz): δ 8.15 (s, 1H), 8.12 (s, 1H), 3.95 (m, 4H), 3.42 (m, 4H), 3.07 (quartet, J=5.9Hz, 4H), 2.79 (s, 3H), 2.17 (quartets, J=7.7Hz, 2H).MS(ESI+)：m/z366.0((M+H)+)。
The calculating of LUMO energy.By Semi-empirical quantum program, MOPAC2009 carries out Energy of molecular orbitals calculating. 52calculate by parameterized diatomic differential overlapping (Neglect of Diatomic Differential Overlap, the NDDO) trial and error method of ignoring of PM6.Use the latent vector tracing routine (Eigenvector Following routine) of acquiescence by the Geometrical optimization of all compounds, until reach self-consistent field (SCF).The LUMO energy of each molecule is by proper value (eV) estimation of the suitable track with EIGEN and the generation of VECTORS keyword.
Bacterial isolates, growth conditions and chemical: Mtb bacterial strain is from American Type Culture Collection (American Type Culture Collection, ATCC, and (the Colorado State University of Colorado State University VA), CSU) obtain, and cultivate in roller bottle (Corning Inc) with Middlebrook7H9 meat soup, in meat soup, be supplemented with 0.2% glycerol, 0.05% tween 80 (Tween-80) and albumin-dextrose-catalase (Difco Laboratories) of 10%.The Middlebrook7H10 agar that is supplemented with the oleic acid-albumin-dextrose-catalase (Difco Laboratories) of 0.2% glycerol and 10% is used for manifesting bacterium colony.Mtb H37Rv grows to logarithmic growth mid-term in the 7H9 meat soup of 37 DEG C.Microbiotic and resazurin are purchased from Sigma (St.Louis, MO), and suspend according to the explanation of manufacturers again.The experiment of carrying out in the institute of tuberculosis of Chicago University of Illinois (TB MIC-ITR) is carried out according to disclosed method. 11，3
The mensuration of antibiotics sensitivity: in order to measure the MIC of compound opposing Mtb, carry out the microplate test based on resazurin.In brief, compound is suspended in DMSO again, and after twice dilution scheme at 10-0.08mg/L scope build-in test.Adding bacterial cell～10 5after colony-forming unit (CFU)/mL, 96 orifice plates are hatched 5 days at 37 DEG C.Add subsequently 0.1% the resazurin of 0.05mL, and at 37 DEG C, hatch in addition 2 days.With Fluoroskan Ascent or Victor3 microplate fluorescent agent (Thermo Scientific, USA) 530nm excite with 590nm transmitting under measure fluorescence.Only contain the hole of compound for detection of the autofluorescence of compound.The lowest concentration of drug that suppresses >=90% growth is considered to MIC.Except fluorescence reading, all MIC values are also scored for determining MIC value by range estimation.The MIC variation of twice is considered to be in the limit of error of test.The ultimate density of the DMSO in porose be 0.625%.These data are shown in table 1-5.
Anti-microbial activity to NRP Mtb cell: each compound is measured with LORA the anti-microbial activity of NRP Mtb cell is as discussed previously. 7in brief, by Mtb H37Rv cell suspension in Middlebrook7H12 meat soup and ultrasonic 15 seconds.By substratum dilution, to obtain the OD of 0.03-0.05 570relative light intensity unit (relative light units, RLU) with every 100 μ L3000-7000.The twice serial dilutions of antiseptic-germicide is prepared in black 96 hole microtiter plates with 100 μ L volumes, and adds the cell suspending liquid of 100 μ L.Microplate substratum is placed in anaerobic condition (O 2< 0.16%) under, use Anoxomat Model WS-8080 (MART Microbiology), carry out three times and vacuumize circulation, and fill 10% H 2, 5% CO 2with surplus be N 2mixture.Anaerobism index strip is placed in the inside of chamber, determines O to estimate 2removal.Plate is hatched 10 days at 37 DEG C, be transferred to subsequently environmental gas condition and (be rich in 5%CO 2air) couveuse in, carry out 28 hours " recovery ".The 11st day (28 hours aerobic recover after), by 100 μ L media transfer to luminous for measuring in white 96 hole microtiter plates.MIC is defined as and for the control group of composite medicine not, produces～90% lowest concentration of drug suppressing.
To the cytotoxicity of Vero cell.Adopt active (Luminescent Cell Viability Assay) the test compounds cytotoxicity to Vero cell that detects of Cell Titer-Glo fluorocyte.Vero cell (Vero ATCC CCL-81) is grown and is maintained in MEM+0.25% foetal calf serum (FBS).Diluted chemical compound liquid is prepared in 96 orifice plates according to the Guide Book of clinical and laboratory standards institute (Clinical and Laboratory Standards Institute, CLSI).The concentration of the compound that each Kong Jun contains 5 μ L and 95 μ L is～5x10 4the host Vero cell of cell/mL.By plate at 37 DEG C at 5% CO 2in hatch 72 hours, subsequently in～30 minutes by plate balance to room temperature.Add subsequently Cell Titer-Glo reagent, and in orbital shaker, mix inclusion 2 minutes and dissolve with inducing cell, and at room temperature further hatch 10 minutes with stabilized illumination signal.Read the reading of plate, and record luminous.Each test panel all contains one group of control group, and described control group only comprises the DMSO control group (for calculating the survival per-cent of all test holes) of substratum (for background deduction) and 0.625%.In addition, each test panel all comprises ATP typical curve, and described typical curve is for calculating the ATP unit of each test hole.Toxicity is defined as the concentration (IC of host cell survival 50% 50).
Mouse lymphoma cell tk+/-→ tk-/-gene mutation test.Experiment is undertaken by the standard program of assessment sudden change potential. 23,24being described in detail in of clone, experimental arrangement and data interpretation is previously open. 54being exposed to 15fa and assessing following end points after 20q in cell: express the cell growth rate of phase, relative suspension growth rate, relatively total growth rate, relative cloning efficiency, mutation frequency and at anti-trifluorothymidine (TFT r) little (diameter≤0.6mm) bacterium colony number in cell.
Solubility test.Compound is worn into fine powder with mortar and pestle, and join in the glass erlenmeyer flask of 25mL, the pH that contains 5mL in flask is 7.4 0.9% normal saline solution.In each flask, add the compound of about 20mg saturated to ensure.Subsequently flask is used to magnetic stirring bar high degree of agitation 48 hours at 25 DEG C.Subsequently by Whatman pvdf membrane (0.45 μ m aperture) injection filter filtered sample.Use subsequently the concentration of compound in HPLC quantitative measurment supernatant liquor in the maximum absorption wave strong point of each compound.Typical curve (the R subsequently each compound being produced 2value > 0.99) carry out integration, to measure the equilibrium solubility of each compound.
The pharmacokinetic of female CD1 mouse.All pharmacokinetics all carry out in AAALAC and OLAW certification authority according to thinking to grind international animal nursing policy (SRI International ' s animal care policies).Program for pharmacokinetic is followed previously described method. 55in brief, the blood plasma pharmacokinetics of selected BTO derivative is measured afterwards giving female CD1 mouse single dose (100mg/kg) by oral cavity feeding.Each time point place of after dosed administration 5 minutes, 15 minutes, 30 minutes and 60 minutes and 2 hours, 4 hours, 6 hours, 8 hours and 24 hours, collects blood from three mouse.Calculate mean value and the standard deviation of the plasma drug level of each time point.Adopt non-compartment model method (non compartmental methods) ( professional, Version5.2, Pharsight Corp, Mountain View, CA) carry out pharmacokinetic analysis.Calculate following parameters: the time (T that reaches maximal plasma concentration maximum), maximal plasma concentration (C maximum), the time of being extrapolated to is the maximal plasma concentration (C0) of 0 o'clock, to the area (AUC under the plasma concentration-time curve of final time point finally) and to the area (AUC under infinitely-great plasma concentration-time curve nothing thoroughly).
The abbreviation using: ATCC, American Type Culture Collection: BTO, 1,2,4-phentriazine, two-N-oxide compound; CC50, cytotoxicity concentration; CDCl 3, deuterochloroform; CD 3oD, deuterated methanol; CFU, colony-forming unit; CLSI, clinical and laboratory standards institute; CSI., Colorado State University; DOTS, short distance DOTS; E1/2, single electron reduction potential; ES-MS, electrospray ionization mass spectrum; FBS, foetal calf serum; LC-MS-MS, Liquid Chromatography-Tandem Mass Spectrometry instrument; LCQ, liquid chromatography quadrupole; LORA, hypoxemia restores test; MDR-TB, multidrug resistance TB; MEM, MEM; MLM, Mouse Liver microsome; MOLY, mouse lymphoma cell mutant test; Mtb, mycobacterium tuberculosis; NRP, non-copy to hold stay; RLU, relative light unit; SD, standard deviation; SI, selectivity index; TB, tuberculosis; TFAA, trifluoroacetic anhydride; TI, therapeutic index; TPZ, Win-59075; XDR, wide spectrum resistance.
The anti-microbial activity spectrum of support information table 1.BTO
The experimental technique of the dull and stereotyped bulk testing of salmonella/microsomes (Ai Musishi screening).
In the case of there is and not existing the rat liver metabolism activation system of the Aroclor1254 induction that contains 10%S9 (MA), the dull and stereotyped hybrid system that employing contains Salmonella typhimurium test strain TA98 and TA100, screens the microbial mutation activity of experimental compound.Confirming, there is suitable hereditary feature in the bacterial strain using in this research.By dissolve each test sample preparation stock solution with 5mg/ml in dimethyl sulfoxide (DMSO) (DMSO).Subsequently, by these initial deposit concentration serial dilutions, (dose volume is 100 μ dosage particles l) to obtain 100,50,10,5,1,0.5 and 0.1 μ g/ flat board.For the condition of revertant counting and background bacterium lawn, by test slab with contrast flat board and compare.When the average number of revertant bacterium colony on test slab exceedes average solvent control batch total number at least when twice, test article is considered to mutagenic compound.In the time of assessment sudden change response, also consider dosage correlation.Cytotoxicity is by some parameter estimations: on test slab revertant colony number object significantly reduce, the formation of the removing of background bacterium lawn growth or disappearance, the non-revertant bacterium colony of point-like, or the disappearance completely of bacterial growth.
The assessment of 15fa and 20q in the dull and stereotyped bulk testing of support information table 2. salmonella/microsomes (Ai Musishi screening).
The data of positive controls:
Without metabolism activation
2NF 2-nitrofluorene
SA sodiumazide
2AN 2-amino anthracene (10%S9)
(10％S9)
DMSO methyl-sulphoxide+S9
(+S9)
The data of 15fa:
Without metabolism activation
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Claims (15)

1. there is compound or its pharmacy acceptable salt or the steric isomer of formula I structure:
Wherein:
Each X is H, halogen, alkyl, OR, SR, NR ' R, BR ' R, heterocycle or another functional group independently, and wherein each R is H, halogen, alkyl or another functional group independently;
W is N, C, O, S, H or B or another connection atom;
Each A and B are H or the optional alkyl replacing, and it can be connected to optionally assorted-cycloalkyl; And
Z is 4-8 ring optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring, at 6,7-, 5, condense on 6-or 7,8-position.
2. the compound of claim 1, wherein:
Each X is H, halogen, alkyl, OR, SR, NR ' R or BR ' R independently, and wherein each R is H, halogen or alkyl independently;
W is N, C, O, S or B;
Each A and B are H or the optional alkyl replacing, and it can be connected to optionally assorted-cycloalkyl; And
Z is 4-8 ring optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring, at 6,7-, 5, condense on 6-or 7,8-position.
3. the compound of claim 1, wherein:
Each X is H, halogen, alkyl, OR, SR, NR ' R or BR ' R independently, and wherein each R is H, halogen or alkyl independently;
W is N, C, O, S or B;
Each A and B are the optional alkyl replacing, and it can be connected to optionally assorted-cycloalkyl; And
Z is 4-8 ring optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring, at 6,7-, 5, condense on 6-or 7,8-position.
4. the compound of claim 1, wherein:
Each X is H, halogen, alkyl, OR, SR, NR ' R or BR ' R independently, and wherein each R is H, halogen or alkyl independently;
W is N;
Each A and B are the optional alkyl replacing, and it can be connected to Heterocyclylalkyl; And
Z is 5 or 6 rings optional existence, saturated or undersaturated, optional replacement, and itself and phentriazine ring condense on 7,8-position.
5. the compound of claim 1, wherein:
Each X is H;
W is N;
A and B are connected to piperidyl or the pyrrolidyl of optional replacement; And
Z is undersaturated 5 rings, and itself and phentriazine ring condense on 7,8-position.
6. the compound of claim 1, described compound is selected from following table:
7. the compound of claim 1, described compound is selected from following table:
8. the compound of claim 1, described compound is selected from:
9. comprise pharmaceutical composition or the test kit of the compound of claim 1 Killing Mycobacterium Tuberculosis (Mtb) medicine different with the second.
10. the method for the compound of preparation claim 3, described method comprises the oxidizing reaction that uses HOF:ACN.
The method that 11. treatment mycobacterium tuberculosis (Mtb) infect, described method comprises: make the people of needs contact the compound of the claim 1 of significant quantity.
The method that 12. treatment mycobacterium tuberculosis (Mtb) infect, described method comprises: make the people of needs contact 1,2 of significant quantity, 4-phentriazine two-N-oxide compound (BTO).
The method of 13. claims 12, it further comprises the subsequent step that detects resulting infection minimizing.
The method of 14. claims 12, it further comprises the preliminary step that detects infection.
The method of 15. claims 12, wherein said infection comprise non-copy to hold stay (NRP) Mtb cell.
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